RNA sequencing of human gut microbiome: adult feces
250 mg of stools from the participants were used for total RNA extraction. For this, the ZR Soil/Fecal RNA MicroPrep kit (Zymo Research) was used following the manufacturers instructions, which not only allows obtaining long RNAs, but also manages to obtain small RNAs. The RNA obtained was quantified by fluorometry using the Quantus Fluorometer (Promega, USA) and analyzed by capillary electrophoresis in Fragment Analyzer (Agilent Technologies, USA). For the qualification of the samples, the RNA should have a RIN equal to or greater than 7.0. Subsequently, the libraries were prepared using the TruSeq Small RNA Library Prep kit (Illumina, USA), following the manufacturer's instructions. The libraries were quantified by fluorometry using the Quantus Fluorometer (Promega, USA) and analyzed by capillary electrophoresis in an Agilent 2100 Bioanalyzer (Agilent Technologies, USA). Finally, sequencing was performed on a HiSeq 50SE (Illumina, USA), according to the manufacturer's instructions.